基于代谢组学和网络药理学的当归四逆汤改善寒凝血瘀致肾损伤作用机制研究  

Mechanism of Danggui Sini Decoction in improving kidney injury caused by blood stasis syndrome based on metabolomics and network pharmacology

作　　者：冯琳琳 唐思琪 农运园 何颖[2] 王倩怡 覃静华 郭玥 苏志恒[1] FENG Lin-lin;TANG Si-qi;NONG Yun-yuan;HE Ying;WANG Qian-yi;QIN Jing-hua;GUO Yue;SU Zhi-heng(Pharmaceutical College,Guangxi Medical University,Nanning 530021,China;First Clinical Medical College,Guangxi Medical University,Nanning 530021,China;Guangxi Institute of Traditional Medical and Pharmaceutical Sciences,Nanning 530015,China)

机构地区：[1]广西医科大学药学院,广西南宁530021 [2]广西医科大学第一临床医学院,广西南宁530021 [3]广西中医药研究院,广西南宁530015

出　　处：《中国中药杂志》2023年第24期6730-6739,共10页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金面上项目(82174263);国家自然科学基金地区科学基金项目(82260922);广西一流学科(药学)建设项目(GXFCDP-PS-2018)。

摘　　要：为了分析当归四逆汤(Danggui Sini Decoction,DSD)改善寒凝血瘀证(blood stasis syndrome,BSS)导致大鼠肾损伤的作用机制。首先,将32只SD雌鼠随机分为4组,正常组和寒凝血瘀组大鼠灌胃等量的蒸馏水,正常+当归四逆汤组以及寒凝血瘀+当归四逆汤组灌胃5.103 g·kg^(-1)当归四逆汤,共计14 d。同时每日给予大鼠冰水浴建立寒凝血瘀模型,并于第14天对寒凝血瘀大鼠皮下注射0.8 mg·kg^(-1)肾上腺素。正常大鼠在37℃下水浴和注射等量的蒸馏水。接着,实验结束后收集大鼠的24 h尿、血清和肾脏,分别进行代谢组学分析、生化指标检测和苏木精-伊红(HE)染色。然后,该研究利用1H-NMR代谢组学技术揭示DSD改善BSS导致大鼠肾损伤调控的代谢网络,结合网络药理学初步阐明DSD起效作用的关键靶点。病理及生化检测结果显示,在DSD干预后肾脏的炎症浸润,血肌酐、血尿素氮和尿蛋白的含量异常得到了显著回调;代谢组学分析发现,DSD降低BSS导致大鼠的肾损伤主要通过回调10个差异代谢物和3条主要代谢通路(牛磺酸与低牛磺酸代谢、柠檬酸循环、乙醛酸和二羧酸代谢)的异常发挥作用;网络药理学分析提示DSD减轻BSS导致大鼠的肾损伤可能与ATP柠檬酸裂解酶(ACLY)和一氧化氮合酶2(NOS2)2个关键基因以及精氨酸生物合成与精氨酸和脯氨酸代谢2条主要代谢通路有关。该研究基于网络调控视角初步揭示了DSD改善BSS导致大鼠肾损伤的作用机制,为进一步深入研究起效过程的分子机制提供了线索和依据。This article analyzed the mechanism of Danggui Sini Decoction(DSD)in improving kidney injury caused by blood stasis syndrome(BSS)in rats.Firstly,32 female SD rats were randomly divided into the following four groups:a normal group and a BSS group,both receiving an equal amount of distilled water by gavage;a normal+DSD group and a BSS+DSD group,both receiving 5.103 g·kg^(-1) DSD orally for a total of 14 days.Daily cold water bath was given to establish the BSS model,and on the 14th day,BSS rats were subcutaneously injected with 0.8 mg·kg^(-1) adrenaline.Normal rats were subjected to the water bath at 37℃and injected with an equal volume of distilled water.After the experiment,24-hour urine,serum,and kidney samples were collected for metabolomic analysis,biochemical measurements,and hematoxylin-eosin(HE)staining.The study then employed 1H-NMR metabolomic technology to reveal the metabolic network regulated by DSD in improving BSS-induced kidney injury and used network pharmacology to preliminarily elucidate the key targets of the effectiveness of DSD.Pathological and biochemical analysis showed that DSD intervention significantly reduced inflammation and abnormal levels of blood creatinine,blood urea nitrogen,and urine protein in the kidneys.Metabolomic analysis indicated that DSD attenuated BSS-induced kidney injury primarily by regulating 10 differential metabolites and three major metabolic pathways(taurine and hypotaurine metabolism,citrate cycle,and acetaldehyde and dicarboxylic acid metabolism).Network pharmacology analysis suggested that the protective effect of DSD against BSS-induced kidney injury might be related to two key genes,ATP citrate lyase(ACLY)and nitric oxide synthase 2(NOS2),and two main metabolic pathways,i.e.,arginine biosynthesis,and arginine and proline metabolism.This study,from the perspective of network regulation,provides initial insights and evidence into the mechanism of DSD in improving kidney injury induced by BSS,offering a basis for further investigation into the molecular mec

关 键 词：肾损伤 当归四逆汤 血瘀证 代谢组学 网络药理学 

分 类 号：R285[医药卫生—中药学]